In vivo evaluation of venular glycocalyx during hemorrhagic shock in rats using intravital microscopy

Filho, Ivo P. Torres; Torres, Luciana Neves; Sondeen, Jill L.; Polykratis, I. Amy; Dubick, Michael A.

doi:10.1016/j.mvr.2012.11.005

Cited by 61 publications

(63 citation statements)

References 39 publications

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“…Approximately 90% of rats from all groups survived the complete protocol. The systemic responses to hemorrhage in this model are described elsewhere (54,59,60). Furthermore, pooled data from 19 sample animals of different groups confirmed that hemorrhage reduced baseline MAP from 100.1 Ϯ 1.7 to 49.7 Ϯ 3.0 mmHg.…”

Section: Resultssupporting

confidence: 57%

“…The intravital microscopy system is described elsewhere (54). It included a microscope (model BX51WI, Olympus), an immersion objective (Olympus ϫ60, numerical aperture 1.00), a tungsten-halogen lamp for transillumination, a metalhalide bulb for epi-illumination, and selective filter block systems for Texas Red (TR) and fluorescein isothiocyanate (FITC) fluorescence.…”

Section: Methodsmentioning

confidence: 99%

“…The cremaster muscle was exteriorized and positioned flat over a thermostatically controlled pedestal (54). During surgery, the muscle was bathed by a warm KrebsHenseleit solution.…”

Section: Methodsmentioning

confidence: 99%

“…Dextrans (Dx) of different molecular weights [Dx70 (70 kDa) and Dx500 (500 kDa)], labeled with TR or FITC, were used to measure the space occupied by the glycocalyx (54). At baseline, TR-Dx70 (10 mg/ml; Molecular Probes) was injected for determination of the intact glycocalyx thickness, and FITC-Dx500 (10 mg/ml; Sigma) was injected after hemorrhageresuscitation (see below).…”

Section: Methodsmentioning

confidence: 99%

“…Paired images of individual microvascular segments using each tracer were recorded using the charge-coupled device camera and transferred to a computer. Precautions were taken to minimize light exposure and excessive or undesirable probe excitation in studied and adjacent microvessels (54).…”

Section: Methodsmentioning

confidence: 99%

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Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids

Filho

Torres

Salgado

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

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107

108

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Torres Filho IP, Torres LN, Salgado C, Dubick MA. Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids. Am J Physiol Heart Circ Physiol 310: H1468 -H1478, 2016. First published April 1, 2016; doi:10.1152/ajpheart.00006.2016.-The endothelial glycocalyx plays an essential role in many physiological functions and is damaged after hemorrhage. Fluid resuscitation may further change the glycocalyx after an initial hemorrhage-induced degradation. Plasma levels of syndecan-1 and heparan sulfate have been used as indirect markers for glycocalyx degradation, but the extent to which these measures are representative of the events in the microcirculation is unknown. Using hemorrhage and a wide range of resuscitation fluids, we studied quantitatively the relationship between plasma biomarkers and changes in microvascular parameters, including glycocalyx thickness. Rats were bled 40% of total blood volume and resuscitated with seven different fluids (fresh whole blood, blood products, and crystalloids). Intravital microscopy was used to estimate glycocalyx thickness in Ͼ270 postcapillary venules from 58 cremaster preparations in 9 animal groups; other microvascular parameters were measured using noninvasive techniques. Systemic physiological parameters and blood chemistry were simultaneously collected. Changes in glycocalyx thickness were negatively correlated with changes in plasma levels of syndecan-1 (r ϭ Ϫ0.937) and heparan sulfate (r ϭ Ϫ0.864). Changes in microvascular permeability were positively correlated with changes in both plasma biomarkers (r ϭ 0.8, P Ͻ 0.05). Syndecan-1 and heparan sulfate were also positively correlated (r ϭ 0.7, P Ͻ 0.05). Except for diameter and permeability, changes in local microcirculatory parameters (red blood cell velocity, blood flow, and wall shear rate) did not correlate with plasma biomarkers or glycocalyx thickness changes. This work provides a quantitative framework supporting plasma syndecan-1 and heparan sulfate as valuable clinical biomarkers of glycocalyx shedding that may be useful in guiding resuscitation strategies following hemorrhage.

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Section: Resultssupporting

confidence: 57%

Section: Methodsmentioning

confidence: 99%

“…The cremaster muscle was exteriorized and positioned flat over a thermostatically controlled pedestal (54). During surgery, the muscle was bathed by a warm KrebsHenseleit solution.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids

Filho

Torres

Salgado

et al. 2016

American Journal of Physiology-Heart and Circulatory Physiology

Self Cite

107

108

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Hemorrhagic Shock and the Microvasculature

Filho

2017

Comprehensive Physiology

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The microvasculature plays a central role in the pathophysiology of hemorrhagic shock and is also involved in arguably all therapeutic attempts to reverse or minimize the adverse consequences of shock. Microvascular studies specific to hemorrhagic shock were reviewed and broadly grouped depending on whether data were obtained on animal or human subjects. Dedicated sections were assigned to microcirculatory changes in specific organs, and major categories of pathophysiological alterations and mechanisms such as oxygen distribution, ischemia, inflammation, glycocalyx changes, vasomotion, endothelial dysfunction, and coagulopathy as well as biomarkers and some therapeutic strategies. Innovative experimental methods were also reviewed for quantitative microcirculatory assessment as it pertains to changes during hemorrhagic shock. The text and figures include representative quantitative microvascular data obtained in various organs and tissues such as skin, muscle, lung, liver, brain, heart, kidney, pancreas, intestines, and mesentery from various species including mice, rats, hamsters, sheep, swine, bats, and humans. Based on reviewed findings, a new integrative conceptual model is presented that includes about 100 systemic and local factors linked to microvessels in hemorrhagic shock. The combination of systemic measures with the understanding of these processes at the microvascular level is fundamental to further develop targeted and personalized interventions that will reduce tissue injury, organ dysfunction, and ultimately mortality due to hemorrhagic shock. Published 2018. Compr Physiol 8:61-101, 2018.

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Microvascular Endothelial Glycocalyx Surface Layer Visualization and Quantification

Alves,

Breslin

2023

Methods in Molecular Biology

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In vivo evaluation of venular glycocalyx during hemorrhagic shock in rats using intravital microscopy

Cited by 61 publications

References 39 publications

Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids

Plasma syndecan-1 and heparan sulfate correlate with microvascular glycocalyx degradation in hemorrhaged rats after different resuscitation fluids

Hemorrhagic Shock and the Microvasculature

Microvascular Endothelial Glycocalyx Surface Layer Visualization and Quantification

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